麝香酮对人源骨髓间充质干细胞成骨分化的促进作用研究  

Study on the promotion effect of muscone on human bone marrow-derived mesenchymal stem cells osteogenic differentiation

作　　者：刘敏婕[1] 刘朋飞[2] 赵雷[3] 

机构地区：[1]暨南大学第二临床医学院.深圳市人民医院手术室,广东深圳518020 [2]吉林大学药学院再生医学系,吉林长春130021 [3]暨南大学第二临床医学院.深圳市人民医院麻醉科,广东深圳518020

出　　处：《中国实验诊断学》2016年第7期1049-1053,共5页Chinese Journal of Laboratory Diagnosis

基　　金：深圳市知识创新计划(JCYJ20140416122812052);深圳市卫生计生系统科研项目(201401010)

摘　　要：目的分析麝香酮对人源骨髓间充质干细胞成骨分化的促进作用,明确骨髓间充质干细胞联合麝香酮在骨损伤治疗方面的应用优势。方法通过密度梯度离心法,从健康志愿者骨髓中分离间充质干细胞,并采用CCK-8检测法以及流式细胞术对骨髓间充质干细胞的增殖能力和表面特异标记进行鉴定,进一步通过实时定量PCR,检测麝香酮处理后的细胞中成骨基因的表达变化,通过碱性磷酸酶染色,评价麝香酮对人源骨髓间充质干细胞成骨分化的促进作用。结果人源骨髓间充质干细胞可在体外正常增殖,同时,骨髓间充质干细胞表面CD44、CD73、CD90的表达均呈阳性,而CD34的表达为阴性,符合正常间充质干细胞的表型,实时定量PCR结果表明,麝香酮处理后,骨髓间充质干细胞中的成骨相关基因ALP、Collagen I、Runx2、Osterix均有一定的上调,而且,在成骨分化过程中,麝香酮也会使细胞碱性磷酸酶染色的程度加深。结论麝香酮对人源骨髓间充质干细胞的成骨分化具有一定的促进作用,骨髓间充质干细胞联合麝香酮的治疗模式,在骨损伤治疗方面具有一定的应用优势。Objective To analyze the promotion effect of muscone on human bone marrow-derived mesenchymal stem cells（BMSCs）osteogenic differentiation and confirm the application advantage of combination of BMSCs and muscone in the therapy of bone injury.Methods Human BMSCs were isolated from healthy volunteer bone marrow with density gradient centrifugation,and cell proliferation ability and characteristics on cell surface were confirmed with CCK-8method and FACS respectively.The osteogenic genes in BMSCs with the treatment of muscone were further analyzed using real-time quantitative PCR（qPCR）.Finally,the promotion effect of muscone on human BMSCs osteogenic differentiation was evaluated though alkaline phosphatase（ALP）staining.Results Human BMSCs could proliferate normally in vitro,and showed a positive expression of CD44,CD73 and CD90,as well as a negative expression of CD34,which coincided with the normal phenotype.The qPCR results indicated that the expression of osteogenic genes,including ALP,Collagen I,Runx2 and Osterix,could be up-regulated in BMSCs with the treatment of muscone,and the treatment also could deepen the ALP staining during osteogenic differentiation.Conclusion Muscone holds the promotion effect on human BMSCs osteogenic differentiation,and the combination of BMSCs and muscone holds the application advantage in the therapy of bone injury.

关 键 词：骨髓间充质干细胞 麝香酮 成骨分化 

分 类 号：R285[医药卫生—中药学]